Dectin-1通过巨噬细胞极化影响STEMI的机制研究

Mechanism of arginase Dectin-1 affecting STEMI through macrophage polarization

目的 探究Dectin-1通过巨噬细胞极化影响STEMI的机制研究。方法 将45只SPF级SD大鼠随机分为假手术组(n=15)、模型组(n=15)、si-Dectin-1组(n=15),模型组与si-Dectin-1组分别通过尾静脉注射NC-质粒和si-Dectin质粒,2周后建立ST段抬高型心肌梗死(ST-Segment Elevation Myocardial Infarction, STEMI)模型,2周后分离心脏组织,qPCR检测心脏组织Dectin-1表达水平,TTC染色检测心肌坏死面积,Tunel染色检测心肌凋亡水平,Masson染色检测心肌纤维化水平,ELISA检测心脏组织炎症因子白细胞介素-1(Interleukin-1,IL-1)、白细胞介素-6(Interleukin-6,IL-6)、白细胞介素-10(Interleukin-10,IL-10)表达水平,Western blot检测心脏组织M1巨噬细胞和M2巨噬细胞标志物诱导型一氧化氮合酶(Inducible nitric oxide synthase, iNOS),CD86,精氨酸酶1(Arginase 1,Arg-1),CD163表达水平。结果 与假手术相比,模型组Dectin表达水平显著升高,si-Dectin-1组Dectin表达水平显著降低;模型组心肌坏死面积增加,细胞凋亡增加,组织纤维化增加,炎症因子IL-1、IL-6、IL-10表达水平升高,与模型组相比,si-Dectin-1组心肌坏死面积降低,细胞凋亡降低,组织纤维化降低,炎症因子IL-1、IL-6、IL-10表达水平降低,差异具有显著的统计学意义(P<0.01);模型组大鼠中iNOS、CD86、Arg-1和CD163的表达水平均显著增加;与模型组比较,si-Dectin-1组大鼠心脏组织中iNOS和CD86蛋白表达水平下调,Arg-1和CD163蛋白表达水平显著上调,差异有统计学意义(P<0.01)。结论 si-Dectin-1可发挥STEMI大鼠的心肌保护作用,其机制与巨噬细胞M2极化的调控相关。

Objective To explore the mechanism of Dectin-1 affecting STEMI through macrophage polarization.Methods 45 SPF SD rats were randomly divided into sham operation group(n=15),model group(n=15) and si-dectin-1 group(n=15).NC plasmid and si-dectin-1 plasmid were injected into caudal vein respectively.The model of ST segment elevation myocardial infarction(STEMI) was established two weeks later.The heart tissue was isolated two weeks later, and the expression level of Dectin-1 was detected by qPCR,TTC staining was used to detect the area of myocardial necrosis, TUNEL staining was used to detect the level of myocardial apoptosis, Masson staining was used to detect the level of myocardial fibrosis, ELISA was used to detect the expression levels of inflammatory factors interleukin-1(IL-1),interleukin-6(IL-6) and interleukin-10(IL-10),and Western blot was used to detect the markers of M1 macrophages and M2 macrophages in myocardial tissue The expression levels of inducible nitric oxide synthase(iNOS),CD86,arginase 1(Arg-1) and CD163.Results Compared with sham operation, the expression level of dectin in model group increased significantly, and that in si-dectin-1 group decreased significantly(P<0.01);Compared with the model group, the area of myocardial necrosis, apoptosis, tissue fibrosis and the expression levels of inflammatory factors IL-1,IL-6 and IL10 in si-dectin-1 group decreased, apoptosis, tissue fibrosis and the expression levels of inflammatory factors IL-1,IL-6 and IL10 in si-dectin-1 group decreased(P<0.01);The expression levels of M1 macrophage markers iNOS and CD86 and M2 macrophage markers Arg-1 and CD163 in myocardial tissue of model group were significantly increased;Compared with the model group, the expression levels of iNOS and CD86 were down-regulated, and Arg-1 and CD163 protein were significantly up-regulated(P<0.01).Conclusion si-Dectin-1 can play a myocardial protective role in STEMI rats, and its mechanism is related to the regulation of M2 polarization of macrophages.